Effect of lupeol on antioxidants and xenobiotic enzymes in N-Butyl-N-(4-hydroxybutyl) nitrosamine induced bladder carcinogenesis in experimental rats.

OBJECTIVE: Urothelial carcinoma of the bladder is a common malignancy ranked 9th with an estimated 356,600 new cases diagnosed annually worldwide. The study showed the protective effects of Lupeol in N-Butyl-N-(4-hydroxybutyl) nitrosamine induced bladder carcinogenesis in in vivo experimental model. Forty male healthy wistar rats were selected randomly divided into four groups. Group I rats served as healthy control. Group II rats were treated with BBN (150 mg/gavage/twice a week) for 8 weeks. Group III rats were treated with BBN + Lupeol [ Lupeol (50 mg/kg bw/day) treatment was started 1 week prior to the BBN treatment, and it was orally administered for 8 weeks]. Group IV rats were treated with Lupeol alone (50 mg/kg bw/day) for 8 weeks. All the experimental rats were maintained and euthanized at 32nd week. Serum and bladder tissues were collected and examined for biochemical parameters, serum markers and histopathological evaluation. Preventive (BBN + Lupeol) group modulates the activity of antioxidant enzymes such as Superoxide dismutase, Catalase, Reduced glutathione, Glutathione Peroxidase, Thiobarbituric acid reactive substances (TBARS) and drug metabolizing enzymes such as Cytochrome P450, Cytochrome b5, NADPH Cytochrome c reductase, NADPH- Quinone Oxidoreductase 1 and Glutathione-S-transferase when compared to BBN treated rats. Serological markers such as Aspartate aminotransferase (AST) and Alanine aminotransferase (ALT) were significantly (P<0.05) decreased in preventive lupeol treated groups. Lupeol supplementation protects BBN induced bladder carcinogenesis in experimental rats by its antioxidant, anti-inflammatory and antiproliferative properties.

Assessment of Histopathological Grade and Ki-67 Expression in Tobacco and Non-tobacco Habitual Buccal Mucosa Cancer.

Although there are various risk factors in the literature, the established primary risk factor for oral cancer is tobacco and betal-nut chewing habits. It is believed that pathogenesis of oral cancer depends on the aetiology. To assess the histopathological grade and Ki-67 expression in tobacco (smoking/smokeless) and non-tobacco (betal nut/pan masala) habitual buccal mucosa cancer. The cross-sectional study was carried out in Regional cancer centre, Tamilnadu. Proliferative marker, Ki-67 expression was determined by immunohistochemistry using biotin-streptavidin method. The study includes 117 buccal mucosa cancer patients (61 male and 56 female). According to WHO grading system, high frequency observed with well differentiated squamous cell carcinoma 48 (41%) followed by moderate 46 (39.3%) and poorly differentiated 23 (19.7%). The cut-off value 50% was used to categorize Ki-67 expression into low and high labelling index (LI); 96 (82%) buccal mucosa cancer and 4 (3.4%) adjacent normal mucosa patients showed high Ki-67 expression. The present study showed highly significant association of histopathological tumor grade and Ki-67 expression by Chi square and paired t test p < 0.05. All the patients were grouped as tobacco 87 (74.4%) and non-tobacco habitual 30 (25.6%) in 3:1, respectively. Further, the risk habits identified with significant differences of tumor grade (p = 0.028) and Ki-67 at p < 0.05. Thus, the study revealed that the nature of cell differentiation and proliferation was strongly related to consumption of carcinogen in both tobacco and non-tobacco form. Therefore, histopathological grade and Ki-67 could be used as a reliable biomarker to understand the biological behaviour of risk habits which might helpful for further treatment therapeutics.

Buccal Mucosa Carcinoma: A Comparative Relative Risk Analysis between Tobacco and Non Tobacco Users.

INTRODUCTION: In South India, buccal mucosa carcinoma is common cancer due to widespread use of tobacco in different form. Recently, areca nut (non tobacco) chewing habits have increased among young adults resulting in high morbidity and mortality. Thus, there is need to understand role of aetiology and risk of disease. AIM: To analyse relative risk of tobacco users and non tobacco users in buccal mucosa carcinoma. MATERIALS AND METHODS: This prospective comparative study was conducted in Regional Cancer Centre, Kanchipuram, between 2013 and 2016. The subjects were divided as tobacco users and non tobacco users in order to evaluate the Relative Risk (RR) in these groups based on demographic and clinical characteristics by Chi-square analysis and Kaplan-Meier (log-rank test) applied for survival difference. All statistics considered at 95 % CI, p<0.05. RESULTS: A total of 117 subjects were included in the study which constituted 87 (74.3%) tobacco users and 30 (25.7%) non tobacco users. In the study, demographic characteristics of gender [RR=0.365 (0.150-0.886), p=0.023] and age groups {RR=2.026 (0.905-4.996), p=0.04} showed significant difference in both study groups. Similarly, clinical characteristics of TNM (tumour node and metastasis) stage {RR=1.57 (0.338-3.31), p=0.024}, nodal status {RR=2.014 (0.412-4.454), p=0.017}, Grade of tumour {RR=1.293 (0.581-2.878), p=0.015}, perineural invasion {RR=2.601 (0.806-5.32), p=0.012} and extracapsular invasion {RR=1.627 (0.533-2.824), p=0.045} showed significant association. The estimated overall survival was 50%; tobacco users (44%) showed lower survival than non tobacco users (57%) with significant difference (p=0.042, p<0.05). CONCLUSION: The study revealed adverse affect of aetiology on survival and showed lower survival of tobacco users than non tobacco users. Hence, the study concluded that the tobacco consumption in different form is more aggressive than non tobacco consumption.

The relationship between histological differentiation and disease recurrence of primary oral squamous cell carcinoma.

BACKGROUND: Although advance techniques were available for diagnosis and prognosis of oral cancer, histopathology was used as major method in clinical routine. Of all oral subsites, buccal mucosa squamous cell carcinoma is aggressive in nature with poor survival. Therefore, the aim of the present study was to evaluate the relation of tumor histopathological grade with disease recurrence of buccal squamous cell mucosa carcinoma. MATERIALS AND METHODS: A retrospective study was carried out in regional cancer research institute, Tamil Nadu. Demographic, histopathological and participant's follow-up details were collected from medical records. RESULTS: Of 198 participants, high frequently encountered with well-differentiated squamous cell carcinoma (n = 98, 49.5%). The clinical characteristics of lymphovascular invasion (P = 0.031), perineural invasion (P = 0.019), tumor stage (P = 0.004), tumor depth (P = 0.048), lymph node (P = 0.02) and metastasis (P = 0.043) had significant association with histopathological grade. In addition, the treatment strategies (P = 0.014) also showed significance at P < 0.05. Further, multivariate revealed cell differentiation (P = 0.048), tumor size (P = 0.037) and depth (P = 0.021) as independent hazard risk of the development of disease using recurrence-free survival of participants at P < 0.05. Of 198 participants, 24 (12.1%) recurrences reported during 34-month follow-up period and the overall estimated recurrence-free survival was 52%. The high frequency of recurrence, 12 (50%), was identified with moderately differentiated tumor cells. However, poorly differentiated tumor showed significantly lower survival (28%) than moderate (54%) and well differentiated (81%) by Kaplan-Meier analysis using log-rank test (P = 0.004, P < 0.05). CONCLUSIONS: The present study concludes high frequency of recurrence observed in moderately differentiated and also revealed lower survival in poorly differentiated tumor. Hence, further treatment plans should focus on moderate and poorly differentiated tumors to improve survival outcome.

Diindolylmethane and Lupeol Modulates Apoptosis and Cell Proliferation in N-Butyl-N-(4-Hydroxybutyl) Nitrosamine Initiated and Dimethylarsinic Acid Promoted rat Bladder Carcinogenesis.

Bladder cancer has been shown to resist programmed cell death with altered expression of both pro-apoptotic and anti-apoptotic proteins. To study is to investigate the apoptotic properties of Diindolylmethane (DIM) and Lupeol on N-Butyl-N-(4-hydroxybutyl) Nitrosamine (BBN) initiated and Dimethylarsinic Acid (DMA) promoted urinary bladder cancer. Sixty male Wistar rats were divided into 6 groups. Group I: Control. Group II: Rats were experimentally developed bladder carcinogenesis with BBN and DMA. Group III and IV: DIM and lupeol were administered after BBN treatment for 28 weeks. Group V and VI: DIM and lupeol alone treatment for 36 weeks. All the experimental rats were maintained and euthanized after 36 weeks protocol. Urinary bladder tissues were collected and processed for further investigations. Apoptotis and cell proliferative marker such as Bax, Bcl-2, caspase-3, caspase-9 and PCNA were quantified using immunohistochemical analysis. The Immunohistochemical expression of Bax, Bcl-2, caspase-3, caspase-9 and PCNA were aberrant in BBN + DMA treated tumor group. Administration of DIM and lupeol inhibited the progression of bladder cancer, induced the expression of apoptotic Bax, caspase-3, caspase-9 and inhibited the expression of anti-apoptotic Bcl-2, PCNA in the urinary bladder of rats. Administration of diindolylmethane and lupeol treatment induces apoptosis and cellular proliferation by its anti-carcinogenic properties. From our results DIM and lupeol would be the agent or adjunct for the treatment of bladder carcinogenesis.

Prevention of N-nitrosodiethylamine-induced hepatocarcinogenesis by S-allylcysteine.

Chemopreventive effect of S-allylcysteine (constituent of garlic) on N-nitrosodiethylamine (NDEA)-induced hepatocarcinogenesis was evaluated in Wistar rats. Significantly decreased lipid peroxidation products (thiobarbituric acid reactive substances-TBARS and lipid hydroperoxides) with increased level of reduced glutathione, increased activities of glutathione S-transferase, and glutathione peroxidase were observed in liver of NDEA-treated rats when compared with control rats. The activities of superoxide dismutase and catalase were significantly decreased in tumor tissue when compared with control. Administration of S-allylcysteine (SAC) showed the inhibition of tumor incidence, modulated the lipid peroxidation, and increased the reduced glutathione, glutathione-dependent enzymes, superoxide dismutase, and catalase in NDEA-induced carcinogenesis. From our results, we speculate that S-allylcysteine mediates its chemopreventive effects by modulating lipid peroxidation, GST stimulation, and by increasing the antioxidants. Hence SAC prevents cells from loss of oxidative capacity in NDEA-induced hepatocarcinogenesis.

S-allylcysteine inhibits circulatory lipid peroxidation and promotes antioxidants in N-nitrosodiethylamine-induced carcinogenesis.

Effects of S-allylcysteine (SAC), an organosulfur compound of garlic, on circulatory lipid peroxidation and antioxidant levels were evaluated in N-nitrosodiethylamine (NEDA)-induced hepatocarcinogenesis in Wistar rats. Significantly elevated thiobarbituric acid reactive substances in the circulation of rats bearing carcinoma indicated the higher levels of lipid peroxidation which was accompanied by significantly decreased levels of antioxidants (beta-carotene, ascorbic acid, alpha-tocopherol, reduced glutathione, glutathione peroxidase, superoxide dismutase and catalase) when compared with controls. Lipid peroxidation has been implicated as a major cause in cancer development. SAC-administered rats showed the inhibition of tumor incidence and lipid peroxidation with simultaneous elevation in antioxidants. We suggest that SAC exerts its chemopreventive effects by decreasing lipid peroxidation and enhancing the levels of antioxidants in NDEA carcinogenesis by reducing the formation of free radicals.